Document Detail

A method for the determination of enzyme mass loading on an electrode surface through radioisotope labelling.
MedLine Citation:
PMID:  12392945     Owner:  NLM     Status:  MEDLINE    
A direct method has been developed for the quantitation of the amount of immobilised enzymes on biosensor surfaces. This quantity is of key importance in establishing the activity, kinetics and optimal immobilisation conditions in the construction of both amperometric and optical biosensors. Recombinant L-lactate dehydrogenase incorporating both a biosynthetically introduced radiolabel, 3H-leucine, and a hexahistidine peptide tag was immobilised on a poly(aniline) composite film and then quantitated by liquid scintillation counting. It was found that enzyme mass loading was proportional to the concentration of LDH in solution, and also depended on the morphology of the composite film. The LDH mass loading on the composite film doubled when a surface cysteine containing variant was used, possibly due to the covalent attachment of the cysteine to the diiminoquinoid rings of the poly(aniline).
Catherine M Halliwell; Evelyne Simon; Chee-Seng Toh; Philip N Bartlett; Anthony E G Cass
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Biosensors & bioelectronics     Volume:  17     ISSN:  0956-5663     ISO Abbreviation:  Biosens Bioelectron     Publication Date:  2002 Dec 
Date Detail:
Created Date:  2002-10-23     Completed Date:  2003-06-18     Revised Date:  2009-07-14    
Medline Journal Info:
Nlm Unique ID:  9001289     Medline TA:  Biosens Bioelectron     Country:  England    
Other Details:
Languages:  eng     Pagination:  965-72     Citation Subset:  IM    
Department of Biological Sciences, Imperial College of Science, Technology and Medicine, Exhibition Road, London SW7 2AY, UK.
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MeSH Terms
Aniline Compounds*
Biosensing Techniques
Coated Materials, Biocompatible / chemistry
Electrochemistry / methods*
Enzyme Activation
Enzymes, Immobilized / analysis*,  chemistry
Equipment Design / methods*
Isotope Labeling / instrumentation,  methods*
L-Lactate Dehydrogenase / analysis*,  chemistry
Leucine / chemistry
Microscopy, Electron, Scanning
Recombinant Proteins / analysis,  chemistry
Scintillation Counting / methods
Sensitivity and Specificity
Reg. No./Substance:
0/Aniline Compounds; 0/Coated Materials, Biocompatible; 0/Enzymes, Immobilized; 0/Recombinant Proteins; 0/polyaniline; 61-90-5/Leucine; EC Dehydrogenase

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